A new immunohistochemical method to evaluate the development of vestibular compensation after unilateral labyrinthectomy in rats.

BACKGROUND: Unilateral labyrinthectomy (UL) causes the disappearance of ipsilateral medial vestibular nuclear (ipsi-MVe) activity and induces spontaneous nystagmus (SN), which disappears during the initial process of vestibular compensation (VC). Ipsi-MVe-activity restores in the late process of VC. OBJECTIVE: We evaluated the late process of VC after UL in rats and examined the effects of thioperamide (H3 antagonist) on VC. MATERIALS AND METHODS: MK801 (NMDA antagonist)-induced Fos-like immunoreactive (-LIR) neurons in contra-MVe, which had been suppressed by NMDA-mediated cerebellar inhibition in UL rats was used as an index. RESULTS: The number of MK801-induced Fos-LIR neurons in contra-MVe gradually decreased to the same level as that of sham-operated rats 14 days after UL. Thioperamide moved the disappearance of the MK801-induced Fos-LIR neurons 2 days earlier. The number of MK801-induced Fos-LIR neurons in thioperamide-treated rats was significantly decreased, compared with that of vehicle rats on days 7 and 12 after UL. But, thioperamide did not influence the decline of SN frequency in UL rats. CONCLUSION: These findings suggested that the number of MK801-induced Fos-LIR neurons in contra-MVe was decreased in concordance with the restoration of ipsi-MVe-activity during the late process of VC after UL and that thioperamide accelerated the late, but not the initial process of VC.

Location and direction of the nutrient artery to the first metatarsal at risk in osteotomy for hallux valgus.

BACKGROUND: Osteotomy for hallux valgus interrupts intraosseous blood supply to the first metatarsal, presumably causing non-union, delayed union, or osteonecrosis of the head of the first metatarsal. We investigated the first metatarsal nutrient artery, arising from the first dorsal metatarsal artery, and identified aspects of surgical technique contributing to nutrient artery injury. METHODS: Enhanced computed tomography scans of 8 feet of 8 fresh cadavers were assessed. Barium was injected through the external iliac artery; location and direction of the first metatarsal nutrient artery was recorded. RESULTS: Mostly, the nutrient artery entered the first metatarsal at the distal third or junction of the middle and distal thirds obliquely from a proximal direction coronally; entry point and direction varied axially. Saw blade overpenetration alone or with extensive capsular stripping might damage the artery. CONCLUSIONS: Location and direction of the first metatarsal nutrient artery was established.

Native rotational knee kinematics are lost in bicruciate-retaining total knee arthroplasty when the tibial component is replaced.

PURPOSE: To compare the kinematics between native knees and knees that have undergone bicruciate-retaining (BCR) total knee arthroplasty (TKA) with cruciate-retaining (CR) TKA converted from BCR TKA in the same whole-body cadaveric specimen using a navigation system and, if differences exist, to investigate the point at which normal kinematics are lost during the procedure. METHODS: The rotational kinematics throughout passive flexion of the native knee and of knees after meniscectomy, femoral replacement, BCR TKA, or CR TKA were assessed in nine fresh frozen cadavers using an image-free navigation system. RESULTS: The rotational kinematic pattern of a knee after BCR TKA was different from that of a native knee, especially in the early flexion phase, and was similar to that after CR TKA. Screw-home movement was not observed after BCR TKA, but still occurred after meniscectomy or femoral replacement with intact cruciate ligaments and an intact tibial articular surface. CONCLUSION: The rotational kinematics of the native knee are not always preserved after BCR TKA. Native rotational kinematics are preserved after meniscectomy and femoral replacement, but are lost after tibial replacement in BCR TKA. Surgeons should pay close attention to maintain the anteroposterior stabilizing function of the ACL in BCR TKA, rather than to restore the native rotational kinematics.

Spinal nerve defects in mouse embryos prenatally exposed to valproic acid.

To examine in detail spinal nerve defects induced by prenatal exposure to valproic acid in mice, pregnant ICR mice were subcutaneously injected with a single dose of 400 mg/kg valproic acid on gestational day 6, 7, 8, or 9, and their embryos were observed on gestational day 10. The whole-mount immunostaining using an anti-neurofilament antibody allowed us to identify spinal nerve defects, such as a loss of bundle, anastomosis among bundles arising from adjacent segment, and a disrupted segmental pattern of the dorsal root ganglia, in valproic acid-exposed embryos. The prevalence of spinal nerve defects was the highest in the embryos exposed to valproic acid on gestational day 8 among the experimental groups. Then, effects of the administration dose of valproic acid on the prevalence of spinal nerve defects were examined on gestational day 10 and found to be dose-dependently increased. It was noteworthy that all embryos exposed to 600 mg/kg of valproic acid on gestational day 8 suffered spinal nerve defects. Folic acid (3 mg/kg/day) supplementation during gestational day 6-10 suppressed the prevalence of valproic acid-induced neural tube defects, which are common malformations in offspring prenatally exposed to valproic acid, but not that of spinal nerve defects. Thus, the spinal nerve defects due to prenatal valproic acid exposure might be induced by mechanisms different from those of neural tube defects. Because spinal nerve defects were predicted to be caused by the disrupted segmental arrangement of the somites and/or that of neural crest cells, which was the origin of the dorsal root ganglia and/or abnormal polarity of the somite, this mouse model with spinal nerve defects at high incidence would be useful to examine the effects of valproic acid on the somitogenesis and morphogenesis of somite-associated structures.

A new concept of transforaminal ventral facetectomy including simultaneous decompression of foraminal and lateral recess stenosis: Technical considerations in a fresh cadaver model and a literature review.

Percutaneous endoscopic surgery for the lumbar spine, which was established in the last decade, requires only an 8-mm skin incision and causes minimal damage to the paravertebral muscles; thus, it is considered to be a minimally invasive technique for spinal surgery. It has been used to perform percutaneous endoscopic discectomy via two main approaches: the TF approach is a posterolateral one through the intervertebral foramen and can be done under local anesthesia; the IL approach is a more traditional one through the interlaminar space and is difficult to perform under local anesthesia. Recently, these techniques have been applied for lumbar spinal stenosis (LSS), the TF method for foraminal stenosis under local anesthesia, and the IL method for central and lateral recess stenosis under general anesthesia. In this study, using a fresh human cadaver model, we performed simultaneous decompression of the lateral recess and foraminal stenosis at L4-5 using the TF approach. Computed tomography confirmed enlargement of the lateral recess and intervertebral foramen. This technique, which can be performed under local anesthesia, should benefit elderly patients with LSS and poor general condition due to multiple comorbidities. Finally, we introduce the concept of percutaneous transforaminal ventral facetectomy using a spinal percutaneous endoscope. J. Med. Invest. 64: 1-6, February, 2017.

Regional difference in sulcal infolding progression correlated with cerebral cortical expansion in cynomolgus monkey fetuses.

The present study aimed to specify the cerebral sulci developed by cortical expansion in cynomolgus monkey fetuses. The degree of sulcal infolding was evaluated by the gyrification index (GI), which was quantified using ex vivo magnetic resonance imaging. The correlation of cortical volume with the sulcal GI was most frequent during embryonic days (EDs) 100 to 120. Interestingly, the high correlation was marked during EDs 140 to 150 in restricted primary sulci in prefrontal, parietotemporal and medial temporal regions. The present results suggest that cortical expansion is involved in gyral demarcation by sulcal infolding, followed by the sulcal infolding progression in phylogenetically-newer cortices.

Influence of Medial Collateral Ligament Release for Internal Rotation of Tibia in Posterior-Stabilized Total Knee Arthroplasty: A Cadaveric Study.

BACKGROUND: Previous studies suggested that changes in kinematics in total knee arthroplasty (TKA) affected satisfaction level. The aim of this cadaveric study was to evaluate the effect of medial collateral ligament (MCL) release by multiple needle puncture on knee rotational kinematics in posterior-stabilized TKA. METHODS: Six fresh, frozen cadaveric knees were included in this study. All TKA procedures were performed with an image-free navigation system using a 10-mm polyethylene insert. Tibial internal rotation was assessed to evaluate intraoperative knee kinematics. Multiple needle puncturing was performed 5, 10, and 15 times for the hard portion of the MCL at 90° knee flexion. Kinematic analysis was performed after every 5 punctures. After performing 15 punctures, a 14-mm polyethylene insert was inserted, and kinematic analysis was performed. RESULTS: The tibial internal rotation angle at maximum knee flexion without multiple needle puncturing was significantly larger (9.42°) than that after 15 punctures (3°). Negative correlation (Pearson r = -0.715, P < .001) between tibial internal rotation angle at maximum knee flexion and frequency of puncture was observed. The tibial internal rotation angle with a 14-mm insert was significantly larger (7.25°) compared with the angle after 15 punctures. CONCLUSION: Tibial internal rotation during knee flexion was reduced by extensive MCL release using multiple needle puncturing and was recovered by increasing of medial tightness. From the point of view of knee kinematics, medial tightness should be allowed to maintain the internal rotation angle of the tibia during knee flexion which might lead to patient satisfaction.

Pulsation and Collimation During Fluoroscopy to Decrease Radiation: A Cadaver Study.

BACKGROUND: Awareness of the harmful effects of long-term low-dose radiation is rising. Many studies have assessed both patient and physician exposure to radiation in association with the use of fluoroscopy in the operating room. However, to our knowledge, previous studies have not assessed, in a detailed fashion, the reduction in radiation exposure that pulsation and collimation provide. METHODS: Seven fresh cadavers were irradiated for 5 minutes with C-arm fluoroscopy with use of standard x-ray and pulsed and collimated x-ray beams. The x-ray sources were placed under the table, over the table, and lateral to the table. Radiation exposure doses were measured at different points, such as the center of the radiation field on the cadaver as well as at the locations of the surgeon's hand and thyroid gland. In addition, Monte Carlo simulation (a physics equation to predict exposure) was performed to estimate the dose reduction and to confirm the experimental results. RESULTS: The radiation exposure doses associated with the use of pulsed fluoroscopy (8 times per second) were reduced by approximately 30% for the patient and by approximately 70% for the surgeon's hand and thyroid gland as compared with those associated with the use of continuous fluoroscopy. The radiation exposure doses associated with the use of collimated beams were reduced to approximately 65% for the surgeon's hand and thyroid gland as compared with those associated with the use of non-collimated fluoroscopy. These results were consistent with the simulation, and the phenomena could be appropriately explained by physics. CONCLUSIONS: The present study revealed the effectiveness of pulsed and collimated x-ray beams in reducing radiation exposure doses resulting from C-arm fluoroscopy. Surgeons should consider using the techniques of pulsed fluoroscopy and collimation to protect patients and themselves from radiation. CLINICAL RELEVANCE: This study presents data regarding the reduction of radiation exposure provided by pulsed fluoroscopy and collimation.

The Pullout Strength of Pedicle Screws Following Redirection After Lateral Wall Breach or End-plate Breach.

STUDY DESIGN: Using fresh cadavers, the biomechanical testing were used to examine the pullout strength of each pedicle screw. OBJECTIVE: The aim of this study was to evaluate pullout strength of (1) a redirected pedicle screw following lateral wall breach; (2) a redirected pedicle screw following end-plate breach; and (3) a pedicle without redirection after end-plate breach without redirection. SUMMARY OF BACKGROUND DATA: Screw malposition, such as lateral wall breach or end-plate breach, is one of the main pitfalls of inserting pedicle screws. METHODS: From 17 fresh spines, 54 vertebrae were harvested. In each vertebra on one pedicle, the screw was inserted correctly down the axis of the pedicle, while on the other pedicle, the screw was inserted to breach the lateral wall or the end-plate. The 18 pedicle screws that breached the lateral wall were then removed and redirected along the correct axis of the pedicle. The 18 pedicle screws that breached the end-plate were removed and redirected along the correct axis of the pedicle. The 18 other pedicle screws that had breached the end-plate were not removed. The pullout force of pedicle screws was measured. RESULTS: First, the mean pullout strength for the redirected screws following lateral wall breach was 24.0% less as compared with the correctly aligned screws. Second, the mean pullout strength for the redirected screws following end-plate breach was 23.3% less as compared with the correctly aligned screws. Third, the mean pullout strength for the pedicle screws end-plate breach was 7.6% less as compared with the correctly aligned screws. CONCLUSION: The pullout strength of redirected pedicle screws after either a lateral pedicle breach or end-plate breach is significantly less than the pullout strength of correctly aligned screw. A pedicle screw that is not redirected after end-plate breach is weaker than a pedicle screw correctly aligned; however, the difference is not significant. LEVEL OF EVIDENCE: N/A.

Radiation Exposure to the Surgeon and Patient During a Fluoroscopic Procedure: How High Is the Exposure Dose? A Cadaveric Study.

STUDY DESIGN: Using fresh cadavers, real-time dosimeters were used to estimate the radiation exposure dose from C-arm fluoroscopy to surgeons, medical staff, and patients during various procedures. OBJECTIVE: The aim of this study was to evaluate the radiation exposure dose from C-arm fluoroscopy, which is used to generate real-time images of the human body, under a variety of conditions and in different areas. SUMMARY OF BACKGROUND DATA: Awareness of the harmful effects of long-term low-dose radiation is rising. There are no all-inclusive reports evaluating the radiation exposure dose to medical staff associated with fluoroscopic procedures that can accurately simulate the real clinical situation. METHODS: Seven fresh cadavers were irradiated for 1, 3, and 5 minutes with C-arm fluoroscopy. The x-ray source was positioned under the table, over the table, and laterally. Radiation exposure doses were measured at different simulated areas such as the center area, and the surgeon's hand or thyroid gland. RESULTS: There were significant differences in the radiation exposure dose under different conditions and for different irradiated areas. The risk of direct and scatter radiation exposure was the greatest with the lateral position, which increased by more than 200 times and more than 30 times, respectively, compared with that from a position under the table. Direct radiation was attenuated to less than one-hundredth after passing through the body of the cadaver. All radiation exposure doses were positively correlated with total exposure time. CONCLUSION: Our study revealed the direct and scatter radiation exposure dose from C-arm fluoroscopy to different areas under a variety of conditions when fluoroscopy is used to generate real-time images of the human body. Our results serve as a guide for medical staff to understand the risk of radiation exposure during each fluoroscopic procedure. Medical staff, especially surgeons, should consider how to protect themselves and reduce radiation exposure by using appropriate shielding. LEVEL OF EVIDENCE: 4.

A 5-HT2A/2C receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane, mitigates developmental neurotoxicity of ethanol to serotonergic neurons.

Prenatal ethanol exposure causes the reduction of serotonergic (5-HTergic) neurons in the midbrain raphe nuclei. In the present study, we examined whether an activation of signaling via 5-HT2A and 5-HT2C receptors during the fetal period is able to prevent the reduction of 5-HTergic neurons induced by prenatal ethanol exposure. Pregnant Sprague-Dawley rats were given a liquid diet containing 2.5 to 5.0% (w/v) ethanol on gestational days (GDs) 10 to 20 (Et). As a pair-fed control, other pregnant rats were fed the same liquid diet except that the ethanol was replaced by isocaloric sucrose (Pf). Each Et and Pf group was subdivided into two groups; one of the groups was treated with 1 mg/kg (i.p.) of 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), an agonist for 5-HT2A/2C receptors, during GDs 13 to 19 (Et-DOI or Pf-DOI), and another was injected with saline vehicle only (Et-Sal or Pf-Sal). Their fetuses were removed by cesarean section on GD 19 or 20, and fetal brains were collected. An immunohistological examination of 5-HTergic neurons in the fetuses on embryonic day 20 using an antibody against tryptophan hydroxylase revealed that the number of 5-HTergic neurons in the midbrain raphe nuclei was significantly reduced in the Et-Sal fetuses compared to that of the Pf-Sal and Pf-DOI fetuses, whereas there were no significant differences between Et-DOI and each Pf control. Thus, we concluded that the reduction of 5-HTergic neurons that resulted in prenatal ethanol exposure could be alleviated by the enhancement of signaling via 5-HT2A/2C receptors during the fetal period.

Recurrence of type I Modic inflammatory changes in the lumbar spine: effectiveness of intradiscal therapy.

Here we report a case of recurrence of Modic type I inflammatory changes in the lumbar spine. A 49-year-old man was referred to our department with a history of chronic low back pain of at least 20 years. At the first consultation, he complained of low back pain only and had no other symptoms such as leg pain, numbness, or weakness. Although his pain was typically mild, he experienced one or two episodes of severe and incapacitating low back pain a year. After two intradiscal steroid injections, his pain disappeared immediately and completely. After 6 months of conservative treatment, Modic type I change switched to Modic type II change. However, 12 months after the first treatment, he once again experienced severe low back pain. Follow-up magnetic resonance imaging (MRI) indicated recurrence of Modic type I change that was stronger than the first occurrence. Two intradiscal injections relieved the pain. Six months after the second episode, follow-up MRI showed another switch of Modic type I change to Modic type II change. Switches of Modic change have been controversial, with mixed findings on pain, natural history, and degenerative changes. The present case reinforces the notion that Modic type I change corresponds to reversible local inflammation.

Prenatal ethanol exposure impairs passive avoidance acquisition and enhances unconditioned freezing in rat offspring.

Previous studies have suggested that ethanol exposure during brain development affects responses to fear and anxiety after maturity. To clarify in detail the impaired behavior related to fear and anxiety seen in rat offspring prenatally exposed to ethanol, their behaviors were observed using an elevated T-maze (ETM) test, which allows assessment of passive avoidance acquisition and one-way escape separately, and an elevated open platform (EOP) test for the assessment of unconditioned freezing against innate fear. The ETM test revealed that acquisition of passive avoidance was significantly inhibited in prenatally ethanol-exposed rats, while their escape behavior was not altered. In the EOP test, the duration of the freezing behavior was significantly elongated in prenatally ethanol-exposed offspring. Thus, we concluded that prenatal ethanol exposure could impair acquisition of passive avoidance, while it could facilitate a response related to unconditioned fears in rat offspring.

Fetal gyrification in cynomolgus monkeys: a concept of developmental stages of gyrification.

Our article summarizes a series of studies about fetal gyrification and its relation to cerebral growth in cynomolgus monkeys. Based on the cerebral growth (i.e., brain weight, cerebral volume, and frontooccipital length of the cerebral hemisphere) and the developmental pattern of gyrification in each sulcus of cynomolgus monkeys, we divided the gyrification process into four stages: Stage 1. Demarcation of cerebral lobes and limbic gyri; Stage 2. Demarcation of neocortical gyri; Stage 3. Emergence of secondary and tertiary sulci; and Stage 4. Growth of sulcal length and depth. Each stage of those gyrification processes was influenced by different developmental events, such as the emergence of corticocortical long-associative fiber tracts, cortical maturations, and subcortical white-matter development. This is the first report to systematically propose gyrification processes closely related to the order of phyologenetical development of the cerebral cortex in primates.

Neuroanatomic and magnetic resonance imaging references for normal development of cerebral sulci of laboratory primate, cynomolgus monkeys (Macaca fascicularis).

Cynomolgus monkey (Macaca fascicularis) is a popular laboratory primate belonging to Old World monkeys, which are the group most closely related to humans except for the apes. This paper summarizes a series of our studies regarding the development of cerebral sulci and gyri in this primate, and the stated possibility of evaluation of the sulcal development for assessing the developmental toxicity testing. The cerebrum of cynomolgus monkeys experienced a regular sequence of emergence of sulci and gyri on gross observation while such timetables corresponded to those obtained by magnetic resonance imaging (MRI) with a lag time of 10-30 days. When the timetables for the emergence of anatomically identical primary sulci and gyri were compared between cynomolgus monkeys and humans, their chronological sequences were comparable, while some sulci and gyri located on the phylogenetically newer cortical region in humans emerged earlier in monkeys. The present paper further indicates brief procedures for evaluating cerebral abnormalities and/or maturity using brain specimens without MRI measurements. The primary sulcal lengths measured by the 'cotton thread' method were a brief index of the degree of regional gyrification. As the development of a calcarine sulcus was closely correlated with morphological maturation of the lateral ventricle, which changed drastically during embryonic days (EDs) 90-100, the cerebral maturity on ED 100 could be evaluated by the infolding of that sulcus. Thus, the present paper provides gross anatomical and MRI references and brief procedures for investigating the normality of the development of cerebral sulci and gyri of laboratory primates, cynomolgus monkeys.

Correlation between formation of the calcarine sulcus and morphological maturation of the lateral ventricle in cynomolgus monkey fetuses.

In the present study developmental changes in the cerebral sulci and volumes of subcortical and archicortical structures of the cerebrum in cynomolgus monkey fetuses were examined with T(1)-weighted magnetic resonance (MR) images in 3D. On the embryonic day (ED) 90, the lateral ventricle had still an immature vesicular shape in the occipital region of the cerebrum, and it dramatically closed its lumen by ED 100. In that period the calcarine sulcus progressively infolded from the medial surface of the cerebral hemisphere narrowing the lumen of the lateral ventricle in the occipital region. Volume of the lateral ventricle decreased in the period ED 90-100, increasing afterwards in spite of increasing volumes of subcortical and archicortical structures such as the caudate nucleus, putamen, globus pallidus, amygdala and hippocampal formation. During the same time, the volume of the germinal matrix around lateral ventricles decreased to disappear completely by ED 120. These results suggest that the morphological maturation of lateral ventricle is linked to the development of calcarine sulcus in cynomolgus monkey fetuses. The degree of infolding of calcarine sulcus on ED 100 would be useful as a gross anatomical landmark for evaluating the cerebral maturation in cynomolgus monkey fetuses.

Sexual dimorphism of sulcal length asymmetry in the cerebrum of adult cynomolgus monkeys (Macaca fascicularis).

The present study aimed to quantitatively clarify the gross anatomical asymmetry and sexual dimorphism of the cerebral hemispheres of cynomolgus monkeys. While the fronto-occipital length of the right and left cerebral hemispheres was not different between sexes, a statistically significant rightward asymmetry was detected in the cerebral width at the perisylvian region in females, but not in males (narrower width of the left side in the females). An asymmetry quotient of the sulcal lengths revealed a rightward asymmetry in the inferior occipital sulcus and a leftward asymmetry in the central and intraparietal sulci in both sexes. However, the laterality of the lengths of other sulci was different for males and females. The arcuate sulcus was directed rightward in males but there was no rightward bias in females. Interestingly, the principle sulcus and lateral fissure were left-lateralized in the males, but right-lateralized in the females. The results suggest that lateralization patterns are regionally and sexually different in the cerebrum of cynomolgus monkeys. The present results provide a reference for quantitatively evaluating the normality of the cerebral cortical morphology in cynomolgus monkeys.

Zebrin II expressing Purkinje cell phenotype-related and -unrelated cerebellar abnormalities in Cav2.1 mutant, rolling mouse Nagoya.

Rolling mouse Nagoya is an ataxic mutant mouse that carries a mutation in a gene encoding for the alpha 1A subunit of the voltage-gated P/Q-type Ca2+ channel (Cav2.1). This report summarizes our studies and others concerning cerebellar abnormalities in rolling mice based on chemical neuroanatomy. While there are no obvious cerebellar deformations in this mutant mouse, the altered functions of Purkinje cells can be revealed as a reduced expression of type 1 ryanodine receptor (RyR1) in all Purkinje cells uniformly throughout the cerebellum, and as an ectopic expression of tyrosine hydroxylase (TH) in the Purkinje cell subsets with the zebrin II-immunopositive phenotype. As the mutated Cav2.1 channel is expressed at uniform levels in all Purkinje cells, its copresence with RyR1 staining suggests that a Cav2.1 channel dysfunction links with the expression of RyR1 in Purkinje cells of rolling mice. However, an ectopic expression of TH in the Purkinje cells is topologically related to the projection of corticotrophin-releasing factor-immunopositive climbing fibers rather than expression of the mutated Cav2.1 channel. On the other hand, increased levels of serotonin (5-HT) in 5-HTergic fibers were revealed immunohistochemically in Purkinje cells of the vermis of rolling cerebellum. Thus, to determine whether or not cerebellar abnormalities are related to Purkinje cell populations revealed by zebrin II expression is essential for enhancing our understanding of the pathogenesis of hereditary cerebellar ataxic mutants such as rolling mice.

Alternating array of tyrosine hydroxylase and heat shock protein 25 immunopositive Purkinje cell stripes in zebrin II-defined transverse zone of the cerebellum of rolling mouse Nagoya.

The present study examined the spatial organization of tyrosine hydroxylase (TH) immunopositive Purkinje cells in the cerebellum of rolling mouse Nagoya with reference to the distribution pattern of the cerebellar compartmentation antigen, heat shock protein 25 (HSP25). Whole-mount immunostaining revealed a striking pattern of parasagittal stripes of TH staining in the rolling mouse cerebellum but not in the control cerebellum. Although the TH stripes resembled the zebrin II stripes in the rolling cerebellum, these two distributions did not completely overlap. The TH stripes were present in the lobules VI and VII (central zone), the lobule X (nodular zone), and the paraflocculus, where zebrin II immunostaining was uniformly expressed. Double immunostaining revealed that TH stripes were aligned in an alternative fashion with HSP25 stripes within the caudal half of lobule VIb, lobules IXb and X, and paraflocculus. Some, but not all, TH stripes shared boundaries with HSP25 stripes. These results revealed an alternating array of TH immunopositive Purkinje cell subsets with HSP25 immunopositive Purkinje cells in the zebrin II-defined transverse zone of the rolling mouse cerebellum. The constitutive expression of HSP25 may prevent the ectopic expression of TH in zebrin II immunopositive Purkinje cell subsets.

Embryonic exposure to ethanol disturbs regulation of mitotic spindle orientation via GABA(A) receptors in neural progenitors in ventricular zone of developing neocortex.

Neural progenitors in the ventricular zone of the developing neocortex divide oriented either parallel or perpendicular to the ventricular surface based on their mitotic spindle orientation. It has been shown that the cleavage plane orientation is developmentally regulated and plays a crucial role in cell fate determination of neural progenitors or the maintenance of the proliferative ventricular zone during neocortical development. We tested if fetal exposure to ethanol, the most widely used psychoactive agent and a potent teratogen that may cause malformation in the central nervous system, alters mitotic cleavage orientation of the neural progenitors at the apical surface of the ventricular zone in the developing neocortex. Fetal exposure to ethanol on E10.5 and 11.5 increased the occurrence frequency of a horizontal cleavage plane that is parallel to the ventricular surface on E 12.5. Administration of picrotoxin, a GABA(A) receptor antagonist, prior to ethanol administration canceled the effect of ethanol with the frequency of horizontal division similar to the control level, although picrotoxin itself did not show any effect on cleavage plane orientation. Phenobarbital, a GABA(A) receptor agonist, induced horizontal cleavage to an extent similar to that induced by ethanol administration. (+)MK801, an antagonist of NMDA receptor that is another major target of ethanol in neural cells, did not affect the cleavage plane of dividing progenitors. These results suggest that fetal ethanol exposure induced alterations in the cleavage plane orientation of neural progenitors in the ventricular zone of the neocortex via the enhancement of the function of GABA(A) receptors.